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Computationally designed ArmRPs (dArmRPs) had to be stabilized by mutations to remove regions of higher flexibility, which were identified by molecular dynamics simulations in explicit solvent.
Primers oNB72 to -81 introduce five point mutations to remove five NdeI restriction sites from the hdrABC operon.
These data are consistent with a model of background selection, because larger LRZs have more potential for deleterious mutations to remove linked polymorphism from the population.
This could be accomplished by introducing synonymous mutations to remove SD-like sequences and use the authors' reporter system to see if there is an enhanced defect in folding or membrane localization.
To introduce silent mutations to remove this homology we utilized alternate codons with a minimum usage of 15% for a given amino acid in predicted T. saccharolyticum open reading frames.
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In the l-threonine biosynthesis pathway, the key enzyme, aspartokinase/homoserine dehydrogenase, encoded by thrA is feedback-regulated and depressed by the product, l-threonine, and a mutation to remove this feedback inhibition has been reported (thrA345).
We used the SOS-response-blocking lexA(Ind−) mutation to remove induction of the SOS response as a complicating factor and found that recA801 could no longer compensate for the loss of recF in the absence of the SOS response.
The protein has an N-terminal hexahistidine tag for nickel chelate affinity purification, the D424A mutation to inactivate the 3′ 5′ exonuclease, the C907S mutation to remove the single native cysteine, and the L744C mutation to provide a unique labeling site on the fingers subdomain.
Our first step to reduce mutations was to remove the possibility of cycling-induced errors by conducting all reactions as single-step extensions.
Another possibility is that by introducing mutations designed to remove its positive charge, we have inadvertently altered some other crucial aspect of the cohesin hinge's function.
The Proteus vulgaris ChABC gene was modified with mutations targeted to remove five cryptic N-glycosylation sites and addition of a mammalian signal sequence and resynthesised with mammalian preferred codons to make a mammalian-compatible engineered ChABC gene (Muir et al., 2010).
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